The evolution and application progress of non-modified drug target discovery CETSA technology / 药学学报

Understanding the research methods for drug protein targets is crucial for the development of new drugs, clinical applications of drugs, drug mechanisms, and the pathogenesis of diseases. Cellular thermal shift assay (CETSA), a target research method without modification, has been widely used since its development. Now, there are various CETSA-based technology combinations, such as mass spectrometry-based cellular thermal shift assay (MS-CETSA), isothermal dose response-cellular thermal shift assay (ITDR-CETSA), amplified luminescent proximity homogeneous assay-cellular thermal shift assay (Alpha-CETSA), etc., which combine their respective advantages and further expand the application scope of CETSA. These technologies are suitable for the entire drug development chain, from drug screening to monitoring the target binding and off-target toxicity of drugs in patients. Based on the author's research experience, this paper reviews the principles of CETSA and related binding technologies, their application in target discovery, and the progress of data processing and analysis in recent years, aiming to provide reference and reference for the further application of CETSA.

Research progress in risk factors of post-transplantation diabetes mellitus / 器官移植

Solid organ transplantation has significantly prolonged the survival of patients with end-stage diseases. However, long-term use of immunosuppressants will increase the risk of post-transplantation diabetes mellitus (PTDM) in the recipients, thereby elevating the risk of infection, cardiovascular disease and death. In recent years, with persistent improvement of diagnostic criteria of PTDM, clinicians have deepened the understanding of this disease. Compared with type 2 diabetes mellitus, PTDM significantly differs in pathophysiological characteristics and clinical progression. Hence, different treatment strategies should be adopted. Early identification of risk factors of organ transplant recipients, early diagnosis and intervention are of significance for improving the quality of life of recipients, prolonging the survival of grafts and reducing the fatality of recipients. Therefore, the diagnosis, incidence and risk factors of PTDM were reviewed in this article, aiming to provide reference for clinicians to deliver prompt diagnosis and intervention for PTDM.

Precise Clinical Positioning of Chinese Patent Medicine in Paradigm of "Target Spectral Effect of Prescription"： A Case Study of Qizhi Tongluo Capsules / 中国实验方剂学杂志

The clinical value of Chinese patent medicine is the core direction of the development of the traditional Chinese medicine industry. The precise clinical positioning determines the way to prove the value of the drug， and is a key link to highlight the clinical value. This paper presented a case study of clinical positioning for Chinese patent medicine， namely Qizhi Tongluo capsules， and the key technical framework of precise clinical positioning of Chinese patent medicine， which was manifested as a comparison of prescription target spectral effect， discovery of core value of prescription， and confirmation of clinical positioning trial. The technical framework was designed to address a range of issues in the realm of precise clinical positioning. Before the clinical positioning trial， based on the multi-component， multi-target， and multi-phenotype data of prescription and clinical indication， the multi-omics network analysis technology was used to identify the core value of the traditional Chinese medicine varieties and predict the potential clinical advantages. Then， based on the predicted clinical advantages， reasonable efficacy indicators were selected， and the clinical efficacy was judged and verified by dynamic and flexible innovative clinical trials to improve the success rate of clinical positioning. This research paradigm integrates "omics technology" with "evidence-based" principles and follows the "precise evidence-based" concept. This research aims to provide a new strategy and method for the precise medication and positioning of Chinese patent medicine with traditional Chinese medicine characteristics after being put into the market and provide more technical thinking for traditional Chinese medicine to move towards precise medicine.

Protective Mechanism of Paeoniflorin on Mice with Ulcerative Colitis Based on AMPK/mTOR Autophagy Pathway / 中国实验方剂学杂志

ObjectiveTo explore the protective mechanism of paeoniflorin on mice with ulcerative colitis （UC） through the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） autophagy pathway. MethodUC mouse model was established by allowing mice freely drink 4% DSS， and 56 BALB/c male mice were randomly divided into model group， AMPK inhibitor group （20 mg·kg-1）， paeoniflorin （50 mg·kg-1） + inhibitor （20 mg·kg-1） group， and high dose （50 mg·kg-1）， medium dose （25 mg·kg-1）， and low dose （12.5 mg·kg-1） paeoniflorin groups. After seven days of drug intervention， the protective effect of paeoniflorin on mice with UC was determined by comparing the body weight， disease activity index （DAI） changes， and Hematoxylin-eosin （HE） staining results. Enzyme linked immunosorbent assay （ELISA） was used to detect the levels of tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） in the serum of mice in each group， and immunofluorescence was utilized to detect microtubule-associated protein 1 light chain 3 （LC3） content in the colon， AMPK， mTOR proteins， and their phosphorylated proteins including p-AMPK and p-mTOR in the colon tissue were detected by Western blot， and the mRNA expression levels of AMPK， mTOR， Beclin1， LC3， and p62 were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the blank group， the model group showed a decrease in body mass， an increase in DAI score， and severe pathological damage to the colon. The levels of inflammatory factors including TNF-α and IL-6 increased in serum （P<0.01）， while the protein levels of LC3 and p-AMPK/AMPK were down-regulated in colon tissue， and those of p-mTOR/mTOR were up-regulated （P<0.01）. The mRNA expression levels of AMPK and LC3 were down-regulated， while the mRNA expression levels of mTOR and p62 were up-regulated （P<0.01）. Compared with the model group and the paeoniflorin + inhibitor group， the mice treated with paeoniflorin showed an increase in body mass， a decrease in DAI score， a reduction in pathological damage to colon tissue， and a reduction in the levels of inflammatory factors of TNF-α and IL-6 in serum （P<0.05）. The protein levels of LC3 and p-AMPK/AMPK in colon tissue were up-regulated， while the protein levels of p-mTOR/mTOR were down-regulated （P<0.01）. The mRNA expression levels of AMPK， Beclin1， and LC3 were up-regulated， while the mRNA expression of mTOR and p62 were down-regulated （P<0.01）. The colon tissue of the inhibitor group was severely damaged， and the trend of various indicators was completely opposite to that of the high dose paeoniflorin group. ConclusionPaeoniflorin can enhance autophagy and reduce inflammatory damage in mice with UC by activating the AMPK/mTOR signaling pathway and thus play a protective role.

Comparison of Effect of Hirudo， Notoginseng Radix et Rhizoma， and Their Combinations on Renal Fibrosis in Rats with Chronic Renal Failure / 中国实验方剂学杂志

ObjectiveTo observe the effects of Hirudo， Notoginseng Radix et Rhizoma， and drug pair on renal pathological morphology and protein phosphatase 2A （PP2A）/adenylate activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） signal pathway in rats with chronic renal failure （CRF）. MethodThe 55 male SD rats were randomly divided into a normal group （n=11） and a modeling group （n=44）. The normal group was fed conventionally， and the modeling group was given 0.25 g·kg-1·d-1 adenine by gavage for 28 days to replicate the CRF model. After successful modeling， rats were randomly divided into model group， Hirudo group （3 g·kg-1·d-1）， Notoginseng Radix et Rhizoma group （3 g·kg-1·d-1）， and Hirudo + Notoginseng Radix et Rhizoma group （3 g·kg-1·d-1）， with 9 rats in each group. The normal group and model group were given a constant volume of normal saline by intragastric administration for 30 days. At the end of the experiment， the levels of serum creatinine （SCr） and urea nitrogen （BUN） in all groups were measured. The renal pathological morphology changes were observed by hematoxylin-eosin （HE） staining， Masson staining， and electron microscopy. The mRNA expressions of PP2A， AMPK， and mTOR were detected by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The protein expression levels of PP2A， AMPK， phosphorylation（p）-AMPK， mTOR， and p-mTOR in renal tissue were detected by Western blot. ResultCompared with the normal group， the renal pathological structure changes were obvious， and the levels of SCr and BUN were significantly increased. The mRNA expression of PP2A， protein expression of PP2A， and p-mTOR/mTOR expression were significantly increased， and the p-AMPK/AMPK was significantly decreased in the model group （P<0.05）. Compared with the model group， the renal pathological morphology changes were significantly improved， and the levels of SCr and BUN were significantly decreased. The mRNA expression of PP2A， protein expression of PP2A， and p-mTOR/mTOR expression in the renal tissue were significantly decreased， and the p-AMPK/AMPK was significantly increased （P<0.05） in all groups after drug intervention. In addition， the effect in the Hirudo+Notoginseng Radix et Rhizoma group was better. The mRNA expression levels of AMPK and mTOR in the renal tissue were not significantly different among the normal group， model group， and other groups. ConclusionThe efficacy of Hirudo and Notoginseng Radix et Rhizoma pairs in improving renal fibrosis in rats with CRF is significantly better than that of the single drug， and its improvement on renal fibrosis in rats with CRF may be related to the regulation of PP2A/AMPK/mTOR signaling pathway.

Danggui Shaoyaosan Regulates Autophagy via AMPK/mTOR/ULK1 Signaling Pathway in Rat Model of Metabolism-associated Fatty Liver Disease / 中国实验方剂学杂志

ObjectiveTo investigate the regulatory effect of Danggui Shaoyaosan on adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like kinase-1 （ULK1） signaling pathway in the rat model of metabolism-associated fatty liver disease （MAFLD）. MethodSixty SD rats were randomized into control， model， western medicine （polyene phosphatidylcholine capsules，0.144 g·kg-1）， and low-， medium-， and high-dose （2.44， 4.88， 9.76 g·kg-1， respectively） Danggui Shaoyaosan groups. After being fed with a high-fat diet for 8 weeks， the rats in each group were administrated with corresponding drugs for 4 weeks. At the end of drug treatment， serum and liver tissue were collected for subsequent determination of related indicators. ResultCompared with the control group， the model group showed increased contents of total cholesterol （TC）， triglyceride （TG）， low-density lipoprotein cholesterol （LDL-C） and activities of alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in the serum， increased contents of TC， TG， and free fatty acids （FFAs） in the liver （P<0.01）， and decreased content of high-density lipoprotein cholesterol （HDL-C） in the serum （P<0.01）. Furthermore， the model group showed down-regulated protein levels of p-AMPK， microtubule-associated protein 1 light chain 3B （LC3B） Ⅱ， Beclin1， and ULK1 （P<0.01） and up-regulated protein levels of p-mTOR and ubiquitin-binding protein p62 in the liver （P<0.01）. The hepatic steatosis was obvious and the NAFLD activity score （NAS） and oil red O staining area increased in the model group， （P<0.05， P<0.01）. Compared with the model group， Danggui Shaoyaosan reduced the contents of TC and TG and the activities of ALT and AST in the serum， lowered the levels of TC， TG， and FFA in the liver， down-regulated the protein levels of p-mTOR and p62 （P<0.01）， elevated the serum HDL-C level， and up-regulated the protein levels of p-AMPK， LCBⅡ， Beclin1， and ULK1 in the liver （P<0.05， P<0.01）. Moreover， it alleviated hepatic steatosis and decreased the NAS and oil red O staining area （P<0.05， P<0.01）. ConclusionDanggui Shaoyaosan has therapeutic effect on MAFLD rats by regulating AMPK/mTOR/ULK1 signaling pathway to enhance autophagy.

Phytotoxic and cytogenotoxic assessment of glyphosate on Lactuca sativa L

Abstract The active ingredient glyphosate is the most commercialized herbicide on the world market due to its capability in eliminating weeds. However, it can harm the development of non-target organisms and threaten environmental quality. This study analyzed the effects of potentially toxic concentrations of glyphosate on germination, growth, cell cycle and genomic stability of Lactuca sativa L., and identified the most sensitive variables for assessing the toxicity of this herbicide to this biomonitor. Seeds of L. sativa were germinated in Petri dishes containing a sheet of filter paper moistened with 5 mL of a concentration of glyphosate (1.34, 3.35, 6.70, 10.05, 13.40 mg L-1). Controls consisted of distilled water (negative) and 3 mg L-1 CuSO4 (positive). Macroscopic and microscopic variables were analyzed. The germination of L. sativa was not affected by the concentrations of glyphosate. Root length and shoot height of the plants and the mitotic index decreased from the lowest concentration tested on. The chromosomal anomaly index and frequency of micronuclei increased by 3.2 and 22 times, respectively, with the presence of the lowest concentration of glyphosate compared to the negative control. The observed phytotoxic and cytogenotoxic effects demonstrate the negative influence that glyphosate has on the development of L. sativa. Root length and microscopic variables showed the highest sensitivity. This study warns of the possible harmful effects that glyphosate can have on non-target organisms and suggests greater control over the use of this herbicide to mitigate its environmental impact.

Resumo O ingrediente ativo glifosato é o herbicida mais comercializado do mercado mundial, pela sua capacidade de eliminar as plantas daninhas. No entanto, ele pode prejudicar o desenvolvimento dos organismos não-alvo e ameaçar a qualidade do ambiente. O estudo teve como objetivo analisar os efeitos de concentrações potencialmente tóxicas de glifosato sobre a germinação, o crescimento, o ciclo celular e a estabilidade genômica de Lactuca sativa L., e identificar as variáveis mais sensíveis para avaliar a toxicidade deste herbicida ao biomonitor. Sementes de L. sativa foram germinadas em placas de Petri contendo uma folha de papel-filtro umedecida com 5 mL das concentrações de glifosato (1,34, 3,35, 6,70, 10,05, 13,40 mg L-1). Os controles consistiram em água destilada (negativo) e 3 mg L-1 de CuSO4 (positivo). Variáveis macroscópicas e microscópicas foram analisadas. A germinação de L. sativa não foi afetada pelas concentrações de glifosato. O comprimento da raiz e a altura da parte aérea das plantas e o índice mitótico reduziram desde a menor concentração testada. O índice de anomalias cromossômicas e a frequência de micronúcleos aumentaram, respectivamente, 3,2 e 22 vezes na presença da menor concentração de glifosato em comparação ao controle negativo. Os efeitos fitotóxicos e citogenotóxicos observados demonstram a interferência negativa do herbicida no desenvolvimento de L. sativa. O comprimento da raiz e as variáveis microscópicas foram as que apresentaram maior sensibilidade. Este estudo alerta sobre os possíveis efeitos prejudiciais que o glifosato pode provocar nos organismos não-alvo, sugerindo um maior controle quanto à utilização deste herbicida, a fim de mitigar o seu impacto ambiental.

Phytotoxic and cytogenotoxic assessment of glyphosate on Lactuca sativa L / Avaliação fitotóxica e citogenotóxica do glifosato em Lactuca sativa L

The active ingredient glyphosate is the most commercialized herbicide on the world market due to its capability in eliminating weeds. However, it can harm the development of non-target organisms and threaten environmental quality. This study analyzed the effects of potentially toxic concentrations of glyphosate on germination, growth, cell cycle and genomic stability of Lactuca sativa L., and identified the most sensitive variables for assessing the toxicity of this herbicide to this biomonitor. Seeds of L. sativa were germinated in Petri dishes containing a sheet of filter paper moistened with 5 mL of a concentration of glyphosate (1.34, 3.35, 6.70, 10.05, 13.40 mg L-1). Controls consisted of distilled water (negative) and 3 mg L-1 CuSO4 (positive). Macroscopic and microscopic variables were analyzed. The germination of L. sativa was not affected by the concentrations of glyphosate. Root length and shoot height of the plants and the mitotic index decreased from the lowest concentration tested on. The chromosomal anomaly index and frequency of micronuclei increased by 3.2 and 22 times, respectively, with the presence of the lowest concentration of glyphosate compared to the negative control. The observed phytotoxic and cytogenotoxic effects demonstrate the negative influence that glyphosate has on the development of L. sativa. Root length and microscopic variables showed the highest sensitivity. This study warns of the possible harmful effects that glyphosate can have on non-target organisms and suggests greater control over the use of this herbicide to mitigate its environmental impact.

O ingrediente ativo glifosato é o herbicida mais comercializado do mercado mundial, pela sua capacidade de eliminar as plantas daninhas. No entanto, ele pode prejudicar o desenvolvimento dos organismos não-alvo e ameaçar a qualidade do ambiente. O estudo teve como objetivo analisar os efeitos de concentrações potencialmente tóxicas de glifosato sobre a germinação, o crescimento, o ciclo celular e a estabilidade genômica de Lactuca sativa L., e identificar as variáveis mais sensíveis para avaliar a toxicidade deste herbicida ao biomonitor. Sementes de L. sativa foram germinadas em placas de Petri contendo uma folha de papel-filtro umedecida com 5 mL das concentrações de glifosato (1,34, 3,35, 6,70, 10,05, 13,40 mg L-1). Os controles consistiram em água destilada (negativo) e 3 mg L-1 de CuSO4 (positivo). Variáveis macroscópicas e microscópicas foram analisadas. A germinação de L. sativa não foi afetada pelas concentrações de glifosato. O comprimento da raiz e a altura da parte aérea das plantas e o índice mitótico reduziram desde a menor concentração testada. O índice de anomalias cromossômicas e a frequência de micronúcleos aumentaram, respectivamente, 3,2 e 22 vezes na presença da menor concentração de glifosato em comparação ao controle negativo. Os efeitos fitotóxicos e citogenotóxicos observados demonstram a interferência negativa do herbicida no desenvolvimento de L. sativa. O comprimento da raiz e as variáveis microscópicas foram as que apresentaram maior sensibilidade. Este estudo alerta sobre os possíveis efeitos prejudiciais que o glifosato pode provocar nos organismos não-alvo, sugerindo um maior controle quanto à utilização deste herbicida, a fim de mitigar o seu impacto ambiental.

EGFR inhibitors: clinical aspects, risk factors and biomarkers for acneiform eruptions and other mucosal and cutaneous adverse effects

Abstract The frequency of the use of drugs that act on the epidermal growth factor receptor (EGFR) is increasing, with the consequent onset of cutaneous toxicity, specifically acneiform eruption. The authors extensively review the topic, focusing on describing how these drugs can affect the skin and its appendages, that is, the pathophysiology that encompasses the cutaneous toxicity related to the use of EGFR inhibitors. In addition, it was possible to list the risk factors that may be associated with adverse effects of these drugs. Based on this recent knowledge, the authors expect to aid in the management of patients who are more vulnerable to toxicity, reduce morbidities, and improve the quality of life of patients undergoing treatment with EGFR inhibitors. Other issues related to the toxicity of EGFR inhibitors, such as the clinical aspects of the acneiform eruption grades, and other different types of cutaneous and mucosal reactions, are also included in the article.

Target product profiles for diagnosis of sepsis: Proposing a new approach for diagnostic innovation

Background & objectives: Sepsis, including neonatal sepsis, remains a prevalent cause of morbidity and mortality in low- and middle-income countries such as India, representing 85 per cent of all sepsis-related deaths globally. Early diagnosis and timely initiation of treatment is challenging due to non-specific clinical manifestations and non-availability of rapid diagnostic tests. There is an urgent need for affordable diagnostics with fast turnaround time catering to the needs of end-users. Target product profiles (TPPs) have been found instrumental in developing ‘fit-for-use’ diagnostics, thus reducing the time taken to facilitate development and improving diagnosis. Hitherto, no such guidance or criteria has been defined for rapid diagnostics for sepsis/neonatal sepsis. We propose an innovative approach for developing the diagnostics for sepsis screening and diagnosis which can be utilized by diagnostic developers in the country. Methods: Three-round Delphi method, including two online surveys and one virtual consultation, was adopted to define criteria for minimum and optimum attributes of TPPs and build consensus on characteristics. Expert panel (n=23) included infectious disease physicians, public health specialists, clinical microbiologists, virologists, researchers/scientists and technology experts/innovators. Results: We present a three-component product profile for sepsis diagnosis, (i) screening with high sensitivity, (ii) detection of aetiological agent, and (iii) profiling of antimicrobial susceptibility/resistance, in adults and neonates with an option of testing different considerations. An agreement of >75 per cent was achieved for all TPP characteristics by Delphi. These TPPs are tailored to the Indian healthcare settings and can also be extrapolated to other resource-constraint and high-disease burden settings.

Analysis of Set Up Margin Required during Delivery of Conformal Radiotherapy in Head and Neck Malignancy Using On Board Imaging: Experience from Tertiary Care Center

Background : Adequate dose to Clinical Target Volume is needed to control tumour and to deliver adequate dose without missing the target, this Clinical Target Volume must be encompassed by two margins for uncertainties; first, Internal margin uncertainties and second, set up margin uncertainty will form Planning Target Volume. Three mm setup error of couch location resulted in 38% decrease of minimum target radiation dose and 42 % increase of minimal Spinal Cord and Parotid Gland radiation dose. Aims and Objectives : Objectives of this retrospective study are, before implementation of high precession radiotherapy technique for Head and Neck Malignancy, we want determine optimal 3-dimensional Clinical Target Volume to planning target volume margin and to assess our setup accuracy in our institute, NRS Medical College & Hospital, Kolkata. Material and Methods : We analyzed retrospectively set up error from 691 set Cone Beam CT images of 94 patients. According to Standard Guidelines Target Volume delineated and for creation Clinical Target Volume to Planning target volume margin, we have used 5-7 mm margin around Clinical Target Volume. Results : In 99% patients’ setup deviation were within 0.5 cm. The population systematic error (?) in in Super Inferior; mediolateral; and anterior posterior direction were 0.13 cm, 0.12 cm and 0.14 cm respectively. The population random error in Super Inferior; mediolateral; and anterior posterior direction were 0.021 cm, 0.022 cm and 0.173 cm respectively. Using van Herk formula Clinical Target Volume to Planning Target Volume margin in Super Inferior; mediolateral; and anterior posterior direction were 0.34, 0.47 and 0.32 cm respectively. Corresponding values with Stroom formula 0.28, 0.40 and 0.26 cm respectively. Conclusions : In our study Set up margin of 5mm all around the CTV to create PTV is found to be safe and adequete

Research Progress of Pseudogenes in Hepatocellular Carcinoma / 肿瘤防治研究

Pseudogenes were initially thought to have no function and were called by aliases, such as "junk genes." With the emergence of large-scale genomics projects and more and more experimental studies, pseudogenes have been shown to play an important role in the occurrence and development of solid tumors, especially playing an important regulatory role in the occurrence and develepment of liver cancer, such as regulating the proliferation, apoptosis, invasion, metastasis, and immunity of liver cancer cells. Recent studies showed that pseudogenes can act as regulators of oncogenes and tumor suppressors in hepatocellular carcinoma (HCC) and can thus serve as prognostic markers and even therapeutic targets for this cancer type. In this review, we systematically summarize the mechanisms and functions of different pseudogenes in HCC and present their future prospects as therapeutic targets.

Research progress on the mechanism of Hippo signaling pathway in organ regeneration / 药学实践杂志

Hippo signal pathway is one of the main signal pathways regulating cell proliferation and apoptosis in multicellular animals, which plays a vital role in the maintenance of tissue homeostasis and the regulation of organ growth. Most mammals have limited regenerative potential, and recent studies have shown that Hippo signal pathway is critical in the regeneration of various tissues and organs. The role of Hippo signaling pathway in organ regeneration and the research progress of related targets were introduced, the mechanism of Hippo signaling pathway promoting regeneration analyzes were aralyzed in this review, which provide theoretical reference for the treatment of diseases related to organ regeneration.

A dosimetric study on off-target isocenter plans for stereotactic body radiotherapy of lung cancer / 中华放射医学与防护杂志

Objective:To investigate the effects of off-target isocenter plans with different off-target distances on the plan quality and delivery accuracy of stereotactic body radiotherapy (SBRT) for lung cancer, aiming to provide a reference for the clinical plan design of SBRT for lung cancer.Methods:For 10 lung cancer patients treated with SBRT, isocenter reference plans were designed by setting the plan isocenters at the mass centers of tumors and 60 off-target isocenter plans by setting the isocenters at distances of 1, 3, 5, 8, and 10 cm from the mass centers of tumors. The dosimetric differences between the off-target isocenter plans and the reference plans. Subsequently was analyzed, under different positional errors (0-5 mm). The gamma pass rates (GPRs) of these plans were measured using the Octavius 4D high-resolution dose verification system, and 240 verifications of these plans were completed. The robustness of the delivery accuracy of the reference plans and off-target isocenter plans were analyzed under different positional errors.Results:The off-target isocenter plans yielded slightly worse dose gradient indices than the isocenter reference plans, but there was no statistically significant differences. With an increase in the off-target distance, the mean lung dose (MLD), V20 of normal lungs, as well as the Dmax of bronchi, showed slight upward trends. Compared with the isocenter reference plans, the MLD of the off-target isocenter plans increased by 0.8%, 0.8%, and 1.9% at off-target distances of 1, 3, and 10 cm, respectively, with statistically significant differences ( z = -2.34 to -1.99, P < 0.05), and the V20 of the off-target isocenter plans increased by 2.0%, 2.5%, and 3.7% at off-target distances of 1, 5, and 10 cm, respectively, with statistically significant differences ( z =-2.11 to -1.99, P < 0.05). In the case of a positional error of up to 5 mm, the GPRs of plans with off-target distances of 5 cm and above decreased by more than 1.0% on average and up to a maximum of 3.5%, showing statistically significant differences ( z = 2.13-2.75, P < 0.05). Conclusions:Compared to the reference plans, the off-target isocenter plans showed slightly lower dosimetric quality and less robust delivery accuracy under different positional errors. Therefore, it is necessary to avoid the plans and treatment with too large off-target distances (≥ 5 cm) as far as possible for SBRT of lung cancer.

Applying an ArcCHECK detector to the dose verification for ultra-long target volumes of cervical cancer / 中华放射医学与防护杂志

Objective:To explore the feasibility of applying an ArcCHECK detector to the dose verification for ultra-long target volumes of cervical cancer.Methods:This study retrospectively selected patients suffering from cervical cancer with ultra-long target volumes (lengths: ≥ 26 cm; 50 cases; the ultra-long target volume group) and conventional target volumes (lengths: < 26 cm; 50 cases; the conventional target volume group). Subsequently, this study designed treatment plans using the Volumetric Modulated Arc Therapy (VMAT) technique and then collected and verified doses using an ArcCHECK detector. The dose detection for the conventional target volume group was performed at the central point of the detector (marked by iso and Short-0 cm). Then, the detector was moved for 5 cm along the bed exit direction (marked by iso 1), followed by the dose verification of the ultra-long target volume group (marked by Long-5 cm) and conventional target volume group (marked by Short-5 cm). The geometric parameters (the length and volume of a target volume), mechanical parameters (machine hop count and the duration of irradiation), and gamma pass rates (GPRs) under different detection conditions of each group were analyzed.Results:The target lengths, target volumes, machine hop counts, and irradiation durations of the ultra-long target group were higher than those of the conventional target group ( t = 2.61-18.56, P < 0.05). For the conventional target group, the GPRs at iso 1 were significantly lower than those at iso ( t = 2.14-8.17, P < 0.05). Meanwhile, the GPRs at iso 1 of the ultra-long target volume group were significantly lower than those of the conventional target volume group ( t = -4.70 to -2.73, P < 0.01). The GPRs of each group met clinical requirements for criteria of both 3%/3 mm and 3%/2 mm. Conclusions:The deviation of the positioning center and the length of the target volume serve as primary factors affecting the dose verification result of cervical cancer. For ultra-long target volumes, dose verification can be performed by moving the positioning center, thus ensuring treatment accuracy for cervical cancer patients.

Application of 3D ResSE-Unet-based intelligent delineation of clinical target volume in postoperative radiotherapy for breast cancer / 中华放射医学与防护杂志

Objective:To evaluate the effectiveness and feasibility of 3D ResSE-Unet-based intelligent delineation of clinical target volume (CTV) in postoperative adjuvant radiotherapy for breast cancer.Methods:A total of 974 cases of breast cancer treated in the Cancer Diagnosis and Treatment Center of the Fourth Affiliated Hospital of Guangxi Medical University from September 2018 to June 2022 were enrolled in this study, including 614 cases receiving total mastectomy and 360 cases treated with breast-conserving surgery. They were divided into a training set, a validation set, and a testing set. The training set consisted of 874 cases and was used to build a model of 3D ResSE-Unet-based intelligent CTV delineation. The validation set comprised 40 cases and was used to evaluate the feasibility and effectiveness of the clinical application of AI-based CTV design in the radiotherapy for breast cancer. The testing set was composed of 60 cases and was used to test the accuracy of intelligent CTV. The Wilcoxon rank test was used to compare the Dice similarity coefficient (DSC), 95% Hausdorff distance (HD95), and average surface distance (ASD) obtained using the intelligent delineation model.Results:The intelligent delineation model showed high precision. The CTV of cases treated with total mastectomy (CTV cw) and the CTV of cases treated with breast-conserving surgery (CTV b) had DSCs greater than 0.80 and greater than 0.88, respectively. Therefore, compared with CTV cw, CTV b had a higher DSC (0.91 ± 0.03 vs.0.83 ± 0.05, t = 7.11, P < 0.05). Both CTV cw and CTV b had lower HD 95 [(7.56 ± 3.42) mm vs.(8.77 ± 5.89) mm] and ASD [(1.85 ± 0.71) mm vs.(1.86 ± 0.83)mm], without statistically significant difference ( P > 0.05). The left/right supraclavicular and infraclavicular CTV (CTV2) had DSCs greater than 0.8. CTV2 also had low average HD95 and ASD, without statistically significant difference ( P > 0.05). Conclusions:The 3D ResSE-Unet-based intelligent CTV delineation has better consistency and feasibility in postoperative adjuvant radiotherapy for breast cancer, especially the CTVs after breast-conserving surgery.

The role of status-related neuronal ensembles in tem-poral lobe epilepsy / 中国药理学与毒理学杂志

OBJECTIVE Epileptic networks are char-acterized as two states,seizures or more prolonged inter-ictal periods.However,cellular mechanisms underlying the contribution of interictal periods to ictal events remain unclear.METHODS Here,we present the procedure for labeling seizure-activated and interictal-activated neuro-nal ensembles in mouse hippocampal kindling model using an enhanced-synaptic-activity-responsive element.This technique is combined with genetically encoded effectors to characterize and manipulate neuronal ensembles recruited by focal seizures(FS-Ens)and interictal periods(IP-Ens)in piriform cortex,a region that plays a key role in seizure generation.RESULTS Ca2+ activities and histo-logical evidence reveal a disjointed correlation between the two ensembles during FS dynamics.Optogenetic acti-vation of FS-Ens promotes further seizure development,while IP-Ens protects against it.Interestingly,both ensem-bles are functionally involved in generalized seizures(GS)due to circuit rearrangement.IP-Ens bidirectionally modulates FS but not GS by controlling coherence with hippocampus.CONCLUSION This study indicates that the interictal state may represent a seizure-preventing environment,and the interictal-activated ensemble may serve as a potential therapeutic target for epilepsy.

Effects of fluoride exposure on autophagy and AMPK/mTOR/ULK1 pathway in NG108-15 cells / 中华地方病学杂志

Objective:To investigate the effects of fluoride exposure on autophagy and the expression levels of adenosine monophosphate activated protein kinase (AMPK), mammalian target of rapamycin (mTOR) and Unc-51-like kinase 1 (ULK1) in mouse neuroblastoma and rat glioma fusion cells (NG108-15 cells).Methods:NG108-15 cells were cultured in vitro and divided into control group (0 mg/L), low fluoride group (20 mg/L), medium fluoride group (40 mg/L) and high fluoride group (80 mg/L) according to the final concentration of sodium fluoride, and the cells were collected after 24 h of treatment for standby. NG108-15 cells autophagy was detected by immunofluorescence/immunocytochemistry (IF/ICC method, the autophagy positive control group was treated with chloroquine phosphate); the mRNA expression levels of AMPK, mTOR and ULK1 in each group were detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR); the protein expression levels of autophagy related protein microtubule-associated protein 1 light chain 3B (LC3B), AMPK, mTOR, ULK1, phosphorylation (p)-AMPK, p-mTOR, p-ULK1 in each group were detected by Western blotting. Results:No autophagosome was detected in the control group, and autophagosomes were detected in all the fluoride groups. The protein expression level of LC3B in the low, medium and high fluoride groups (1.80 ± 0.59, 2.16 ± 0.60, 2.30 ± 0.57) was significantly higher than that in the control group (1.00 ± 0.29, P < 0.05). The results of qRT-PCR showed that compared with the control group, the mRNA expression levels of AMPK in medium and high fluoride groups were higher (2.30 ± 0.57, 4.41 ± 1.05 vs 1.00 ± 0.01, P < 0.05); the mRNA expression levels of mTOR in the low, medium and high fluoride groups were lower (0.79 ± 0.04, 0.76 ± 0.09, 0.64 ± 0.10 vs 1.00 ± 0.01, P < 0.05), and the mRNA expression levels of ULK1 were higher (1.81 ± 0.39, 1.96 ± 0.35, 4.22 ± 1.03 vs 1.00 ± 0.01, P < 0.05). The results of Western blotting showed that compared with the control group, the protein expression levels of AMPK (1.21 ± 0.05, 1.20 ± 0.04, 1.30 ± 0.07 vs 1.00 ± 0.03), p-AMPK (1.12 ± 0.05, 1.20 ± 0.06, 1.49 ± 0.07 vs 1.00 ± 0.02), ULK1 (1.16 ± 0.05, 1.26 ± 0.05, 1.15 ± 0.05 vs 1.00 ± 0.04) and p-ULK1 (1.19 ± 0.04, 1.17 ± 0.02, 1.24 ± 0.05 vs 1.00 ± 0.05) in the low, medium and high fluoride groups were higher ( P < 0.05), and the protein expression levels of mTOR were lower (0.77 ± 0.03, 0.60 ± 0.03, 0.55 ± 0.04 vs 1.00 ± 0.04, P < 0.05); the protein expression levels of p-mTOR in the medium and high fluoride groups were lower (0.93 ± 0.05, 0.48 ± 0.02 vs 1.00 ± 0.02, P < 0.05). Conclusion:Fluoride exposure can induce autophagy in NG108-15 cells, and the expression of AMPK and ULK1 are up-regulated, while the expression of mTOR is down-regulated.

Bioinformatic analysis of the expression of KIF14 and its clinical significance and biological functions in clear cell renal cell carcinoma / 现代泌尿外科杂志

【Objective】 To investigate the expression of Kinesin family member 14 (KIF14), and its correlation with clinical prognosis and immune cell infiltration of clear cell renal cell carcinoma (ccRCC). 【Methods】 The correlation between KIF14 expression in ccRCC and different clinicopathological features were analyzed with TCGA, GEO and Ualcan databases. The correlation between KIF14 expression and prognosis was analzyed with Kaplan-Meier method. The correlation between KIF14 expression and immune cell infiltration was analzyed with TIMER. The protein-protein interaction network of KIF14 was conducted with Genemania. The co-expression genes of KIF14 in TCGA-KIRC were picked out in Linkedomics database and were used to perform GO annotations and KEGG pathway enrichment analysis with R software. The biological functions of KIF14 were verified with in vitro functional assay. 【Results】 KIF14 was highly expressed in ccRCC tissue and was positively correlated with clinical stage, pathological grade, and lymphatic metastasis, but negatively correlated with clinical prognosis. KIF14 expression was an independent risk factor for overall survival of ccRCC patients. GO annotations showed that KIF14 was involved in DNA replication, nuclear division, organelle fission, and cell adhesion. KEGG pathway enrichment analysis showed that KIF14 participated in cell cycle and p53 signaling pathway. Genemania analysis indicated KIF14 interacted with CENPE, CIT, KIF23, and other proteins. Timer showed that KIF14 was positively correlated with immune cell infiltration. Knockdown of KIF14 expression suppressed cell proliferation, migration, and invasion of ccRCC. 【Conclusions】 KIF14 may serve as a novel prognostic marker and a potential therapeutic target of clear cell renal cell carcinoma.

Research progress in drugs targeting tumor associated macrophage / 药学学报

Tumor brings great threat to human public health. In recent years, incidence rate and mortality of tumor were rapidly increased in the world. Anti-tumor therapies have undergone the development of cytotoxic therapy, targeted therapy, and immunotherapy. Among them, tumor immunotherapy is rapidly developed and becomes an important anti-tumor therapy in recent years, although it also brings some related side effects. Tumor microenvironment (TME) is composed of immune cells, vascular vessels, fibroblasts, the extracellular matrix, etc. TME significantly affects the efficacy of immunotherapy. Macrophages in the TME are named as tumor associated macrophages (TAMs). Recently, increasing studies have shown that TAMs play an important role in the regulation of tumor immunity, especially in tumor immune surveillance and immune escape. Currently, more and more anti-tumor immunotherapy strategies targeting TAMs are at the development stage. Based on the important role of TAMs in the TME and their potential as therapeutic targets in tumor immunotherapy, we first reviewed the subtypes and functions of TAMs, as well as the roles of TAMs in tumors. Furthermore, we summarized the research progress on anti-tumor strategies targeting TAMs and the current status of drug targeting TAMs. The current review will provide new ideas and novel insights for tumor immunotherapy.